This article isThis article by protected copyright. All rightsreserved. 10.1111/mec.14069doi: differences to lead between the of thisversion c and Version Record.Please copyediting, paginationbeen throughthe andproofreadingtypesetting, process,may which This article acceptedhas been for publication andundergone fullpeer review buthasnot E Fax: 28006, Madrid National Museum Borja Milá Corresponding authors pyrrhocorax Keywords: 1 28006 Madrid ¶ Real, Portugal ofUniversity Trás § Real, Spain ‡ L lote † dePrados,Quinta Sciencesand Biological of (CITAB),University Trás * CARVALHODIOGO CABRAL FRANCISCO MORINHA E type Article :OriginalArticle DateAccepted 09 : Revised Date:08 DateReceived 18 : MR. BORJAID:0000 MILÁ(Orcid
National Museum of Natural Sciences Natural of Museum National Department ofGenetics ofLife andBiotechnology, School andEnvironmental Sciences deInstituto Investigación enRecursosCineg M Laboratory of Ecology Applied These authors contributed equally tothis - xtreme genetic stru Acceptedmail: Article orinha (+34) 915645078(+34) - 2, Lj B5, 5000 dispersal capacity [email protected]
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, This article isThis article by protected copyright. All rightsreserved. and environmental barriers which genetic populations diversity, genetic isolation populations, sothat qualitativelyoverall similar among andregularlysharedbyindividuals regions ofdifferent Genetic are individuals structure genetic chough, asocial c divergence majorunless geographic barriers or habitat cetaceans, among populations Social Abstract Running title E Fax: 28006, Madrid National Museum Sciences Research Natural (MNCN), SpanishNational Council (CSIC), of Guillermo Blanco olonization or -
Accepted Articlemail: [email protected] common (+34) 915645078(+34)
may barriers structure differentiation by adaptation
apes have driven . over 30yearsover revealed , yet recruitment, yet populations is breeding infrequent into and ph highly We that propose :
have
Extreme structure genetic in the Spain adaptation and , gregarious
in birds composed ofatleast 15distinct units.genetic suggesting
in species within species highcognitive humans genetic structure been shownto
. is weakly related
this this Mo
through
limitation . reover, reover,
. Inbirds
r corvid with high dispersal capacity, dispersalcapacity, corvid with high emarkable a limited for role social mechanisms
a patternof by isolation
that most population nuclei showedmost populationnuclei reduce geneflow and . , is unlikely is unlikely to bedue W high dispersalcapacity tois thought long e population divergence to report that report
geograp - distance movementshundreds ofkilometres over genetic chough may capacity hic hic Iberian populationsIberian of the underlie this unprecedented this underlie level of drift induce distance
contribute to to contribute
social solely
and complex in
significantly isolation patternsisolation
in the absence of geographic in the absenceofgeographic M and habitat and habitat usedare types barriers to isolation bydistance or show onitoring relatively relatively
a genetic structure
striking societies not prevent population prevent differentiating of yet yet high levels of
marked red
by
described degree of , such as - i billed disper lopatric. sal , ,
This article isThis article by protected copyright. All rightsreserved. localscales structure at is somethatsocio evidence (Marzluffbehaviour & Among highcognitivecapaci with birds combinationor a of rare,andgenerallyarisesfromrelatively major capacity,barriers, low dispersal geographical al ability tocrossassociated geographical barriers(Pilot several (Orsini isolation withconsistent animals social other than mammals 2013). populations (Ségurel lifestyles socio example ofcomplex ge gene flowamong Slatkin primarily Introduction
Accepted. 2014 Article - cultural traits suchasethnolinguisticboundaries, social organization This mechanism of isolation, mediated been barriers, has bysocial
There isevidenceThere thatcomplex socialprocesses differentiation Genetic non alet 1987). ;
, by colonization (geographic barriers) Foote
by geographic barriers to gene bydistanceWr flowandisolation (e.g. act asbarriers flowandtherebyto gene differentiation promote genetic among - . 2013 human
However, c patterns of isolation by limitation distance), todispersal (i.e.,geographic et alet populations ( populations ;
Andrews primate andcetaceanspeciesprimate withvariable dispersal capa . 2016 both et alet
ne Angell 2005;Angell Clayton when combined withlow dispersal and small size population
( - . 2008; Francisco Francisco omplex patterns social ). culture culture interactions (Laland -
cultural mayfactors played have arole in driving genetic In
2014
among large populations at geographicscalesis Ross 2001 terrestrial Novembre ). , where ,
et al et ty, developedcorvids have complex patterns o . 2007; birds, ). The evolutionary history ofhumans). Theevolutionaryhistory the is
patterns of et al et & Emery ,
or however, Naka . 2009; are also knownare also to isolation isolation
et al et al et neutral 2008
, 2007; Loretto 2007; Loretto
Tishkoff similar tothose
fine . 2010; Kopp . 2012; by (ecologicalbarriers) adaptation ; Gintis
- genetic structure genetic scale genetic structurescale genetic is et al et
Bertrand
play a play role 2011), wherediverse et et al . 2009;Ross et al et in
rarely reported rarely in . 2012), and there . 2012),and there
, humans,
et al et and traditiona . 2014;Kopps city and
in caused are are typically ight 1943;ight . 2014). restricting restricting et et al occur in occur an clearest f social f social . l et et
This article isThis article by protected copyright. All rightsreserved. inlandrocky andcoastal area where areacomprisesIberianThe study the Peninsula in ( widespread corvids L andMaterial methods 30 years from aregion microsatellite loci differentiation. versus explore ( breeding recruitment within among and barriers scales al scalesgeographic (Omland (Abdelkrim Cramp Pyrrhocorax pyrrhocorax ong . 2012
Accepted caves, humancrevices and Article - term term
requires numerous potential , Red Here
the relative rolesthe factors of(e.g. extrinsic geographic distance, orhabitat)barriers, ). 1988 to gene flow,
showing strict mate andnesting to assess dispersalcapacity and recruitment probability Demonstr population - at at billed c , et al et
) we - . wide long continental scales
rul The We infer
intrinsic (e.g. factors . 2012) examine fragmented
ing and sequence y houghs ating
forag
out and monitoring
. - r
However, this has has However, this termmonitoring program ed
alternative alternative the role ofsocialfactorsthe indriving divergencepopulation at large patterns ofgenetic patterns population small e ), a highly), a social
(choughs hereafter)
et al et mostly landscapes(Blanco inopen population structure population structure from s - population made structures
( Fig. - s , but atregionalscales, rare . 2000; scale habitatspecialization scale
from twomitochondrial DNA regions
and sampling genetic 1) factors factors such as limiteddispersal natal natal (Blanco 2003;(Blanco Cuevas&
McCormack
Iberian populationsIberian of thered - nuclei site fidelity dispersal, socialbehaviour) in population driving not corvid
(Blanco
, are been case the are highly social, medium the for European species, stronghold this
distributed across with high capacitywith high dispersal of markedof red
et al et (Banda &Blanco individual genotypes 11 individual at et alet
structure, structure, . 2008;
. 199
through local adaptation in
in mostin at large studies
Europe, et al et
Blanco 6 into breedinginto populations van Els - ; Banda & billed choughs over individual dispersal and . 1996 and
open, capacity -
billed chough
2014). Africa, and Asia
. 2009).
We also et al - sized, long sized, montane and
Blanco 200 . 2012; , geographic They are
.
1998) and
We aim used .
Zhang -
lived the last the data data
9 ed ) nest nest . .
to et et
This article isThis article by protected copyright. All rightsreserved. Accepted precisionlikelihood & Wang screened for minimize biasTo due potential to family from individuals sampling, were sampling each site was negligible for lociexceptPpyA1.9, all whichwasexcluded fromsubsequent analyses. all (95%)intervals derivedfrom CarloMonte 10,000simulations. ofnull Theprobability alleles v.2.2.3 (VanOos null genotyping errors, alleles, largealleledropout stuttering with and from lociexcluded within thedataset.All eachpopulation were forpresence screened the of ampl choughs individuals were Microsatellite Article ring with spotting scopes flocks originnatal 1999; &Tella (Blanco Banda& capturing b periodstudy breeders weregularly searchedbysurveying forringed nesti monitor individually throughout and record presence the ofpreviously ification andgenotyping
were Genomic DNAwas extracted B
dispersal dispersal (Dávila iological samplesiological mo for 2010) reeding pairs the also also marked close close genotyping , andapplying Iberian distribution distribution Iberian range ( regularly et al terhout terhout genotyped at11polymorphic microsatellite loci developedspecifically for movements recruitment andbreeding genetic relatedness (presence of full , . M
nestlings andfull nestlings and . 2015) ultiple capturesultiple ofcommunally roosting werealsoconductedto birds .
et al et
no We
monitored
sibship
( . 2004) usingBonferroni (Dunn ) record the full see Supplementarysee for Methods . Individuals withmissing atthree or data more were loci lecular wereanalysis lecular collected
ed prior,
- to recordthe from feather and samples blood likelihood method -
grown individuals if theywere
Table Table assuming a marked
Blanco 2014). 1 ; movements individuals ( Fig. ringed ringed monogamous -
sibs) using sibs) 1 ( , T ) of long length from order 1985to2016 in . Intwelve ofthes able able as nestlings C - ommunal roosts Sidak) adjustedSidak) confidence details onDNAdetails extraction, of 2 T ) able at . ringed ringed
Throughout COLONY
25 population mating system andan .
2 A MICRO to ). run
individuals total of642total determin ng sites andng sites
, e areas v.2.0.5.8 (Jones (Jones v.2.0.5.8 medium -
CHECKER and foraging the entire the nuclei nuclei e their e their ,
we also we also identified
to
This article isThis article by protected copyright. All rightsreserved. ( statistical parsimony using program networkwasconstructed the diversity haplotypes; KX024343 were submitted toGenBank un Supplementary Methods details) for amplification 2 Mitochondrial DNA was obtainedusing 2008). v.4.4.2 (Rousset usingobtained ( characteri iterations/batch) 2008) Weinberg equilibrium p of individual selection at loci. data setusedinthestudy. genotype one member of full each inbreeding model Bandelt Bandelt H ( O AcceptedND2 Article
and
using using method Markov the chain (10 Partial sequences ofthe Partial disequilibriumLinkage ofloci (LD)between pairs anddeviations from )
H ) were obtainedwith) were were used to assess to were used mitochondrial genetic diversity et et al s E ed number bythe locus per of( alleles - ) calculated calculated with ) KX024378 h , haplotype diversity;, haplotype . 1999 FSTAT , and a s
for CERVUS ). sequencing
isolated populationsisolated &Waller(Keller
v.2.9.3.2,
( er sampling wereer location calculated with equential Bonferroniequential correction - ND2 Polymorphism information(PIC) content population forall loci per sib pair detectedsib pair witha probability >
v.3.0.3 GENAIEX
sequences D
der accession der
control regioncontrol NA indexand fixation ( and analysis SP
K (Kalinowski LOSITAN , average number, average ofdifferences
v.5.10.01 (Librado v.5.10.01 (Librado & Rozas v.6.5 (Peakall & Smousev.6.5 (Peakall ). M , 000 dememorisation tDNA genetic diversitytDNA genetic s
(Antao (Antao
KX024396 (CR)
et al et N . A The sequences of all haplotypesThe sequencesofall identified F ), observed andexpectedheterozygosities and NADH dehydrogenase and NADH dehydrogenase et alet . 2007 IS ) values estimated) values using .
. 2008) was usedto Microsatellite geneticdiversity was -
KX024431 ).
2002
and structure (see
0.90 was included in 0.90 wasincluded in the final
2012) steps, 1000steps, batches and 5000 ) N .
GENEPOP 2009). A ETWORK After runs fourlong indices ; andπ,nucleotide
, ( CR allelic richness (
sequences
v.4.4.2 (Rousset v.4.4.2 (Rousset ( v.4.6.1.3 median H detect evidence detect evidence , number of Hardy GENEPOP gene - joining ) and subunit - A ,
only R ) This article isThis article by protected copyright. All rightsreserved. geographic each provenance within sampling region. of zero assuming individualsuncertainty wassetto coordinates the havethesame that number there model waschosensince true spatialfrequency model,a model allele andafalse null model. Theuncorrelated allele of number ofnuclei of590of process Poisson (equal to thenumber individuals dataset), maximum of inthe a following conditions: 1000 000MCMCinteractions athinningof100,maximum with rate wereinferences carriedout using modelexplicit thatassumes locationofthe study, geographical samples. this the the In for theinferenceofnumber which ofpopulations, implemented canbe aspatial using and implemented in reliabilithe to lead incongruent results among packages ( the under assumptionHardy of clustering algorithms clusteringBayesian analysis Accepted Article Pearse & STRUCTURE GENELAND Red of populations ( of populations differ in slightly the
Crandall 2004; - ty genetic structure, of thepopulation billed
GENELAND v.2.3.4 (Pritchard (Pritchard v.2.3.4
chough population chough population uses a a uses
1770 (3xmaximum ofPoisson process), rate , which Guillot Guillot Reversible Ju Reversible Manel
v.4.0.5 (
use multilocus genotype datato individuals assign to clusters and population genetic differentiation is model – et al et Weinberg andlinkage equilibrium population each within
evidence thatthecorrelated model et alet
20 independent with runs
different methods (François & et al Guillot Guillot . 2005 assumptions principles, andmethodological whichmay . structure wasinferred across Iberia Bayesian using
2005). . 2000) mp Chain Carlo Markov Monte (RJMCMC)algorithm b et al et ; Cullingham &Moehrenschlager The available The available , respectively . 2005a we used
),
BAPS
three different strategies . K Bayesian clustering
varying from varying 1to10underthe
v.6.0 (Corander v.6.0 (Corander
Durand 2010). Tosupport an
may uncorrelated allele overestimate the
2013 et al software ). The . 2008) ly
This article isThis article by protected copyright. All rightsreserved. Accepted approach equal + 1is to mode); patterns betweenruns(at clustering 60%of the20runs least distribution reached a maximum plateau ordecrease; beganto (2)high stability and of The optimal distributionslikelihood were analysed forthreshold scores similarity of0.90 outputs 2009 sam using performed were simulations of 100 for all to select the best Articlewe followed chainMarkov Carlo Monte (MCMC) algorithm. individ parameters: 1000interactions,2000referenceindividuals and 10 interactions forreference estimate admixture coefficientsamong clusters, the program wasrunwiththefollowing clusteringofspatial gr andspatialclustering of “groupspatial “individuals”or posterior modeinfer the (Corander structure ofthegenetic ). , K 000 The uals. The program In and to obtain assignment pr
values (va to finedetect BAPS iterations CLUMPAK server CLUMPAK(Kopelman server
(3)
K a strategy similara strategy approach toan previously reported
K , value value K
the Bayesian clustering Bayesian modelthe u max). max +1nolonger identify new K
rying fromwith each 1to10), 1 (Tishkoff , under amodel ofadm STRUCTURE was selected considering thewas selected followingassumptions: (1) oups was implemented 10replicatesforeach with - STRUCTURE scale population structure et al. et obabilities
infers
analyses were performed also analyses a hierarchical using 2009). )
with
pling localities as priors ( priors as localities pling and to displayand to the inagraphical results interface. the optimal ( numberofpopulations
Thus, theanalysisconsis of individuals et al et S ixture andcorrelatedallelefrequencies. The ixture TRUCTURE TRUCTURE . 2015)wasused ses a ses a stochastic optimization algorithm to
Considering thecomplexityConsidering ofour dataset clusters (i.e.
( Evanno , 000 s , H 000 MCMCit
( of individuals”. In this approach,of Inthis the individuals”. ARVESTER
q ) to cluster each et et al et al
were similar in ,
the geneticstructureat the . 2006),allowing non the .
locprior 2005 to process structure the
for ted of20ted
(Earl & ) highly structured data erations erations after a burn . Thus, population K option) (Hubisz option)
(from To 2to10). (using a K
von Holdt 2012). independent runs independent runs the primarythe ) the likelihoodthe
using a using MCL K max
- The et al et - , in
. This article isThis article by protected copyright. All rightsreserved. AcceptedSupplementary Methods for detailed a in implemented model ratio bottl genetic for evidence the Additionally, (Piry v.1.2.02 flow) eve demographicidentify to data important provides (mutation for Testing departures from mutation permutations thesignificance toassess ofpairwise was chough populations calculated in significance to test at (maximum numberofpermutations by allowed Article ofThe calculation GENA Cockerham1984) markers, andpairwise genetic global differentiation wascharacteri from microsatellite loci 10 andtwo mtDNA ( markers considering numbern the ofsampling procedures described abovefor first level, the separately differentiatedat firstclusters the of level L hc may which EX h aayi o dprue fo nurl qiiru associated equilibrium neutral from departures of analysis The of The level differentiationgenetic among wasestimated populations basedondata
- v.6.5 using the v.6.5 using
rf dsqiiru) r ouain rcse (migration processes population or disequilibrium) drift in level asecond t al et and Jost’s and Jost’s D have influenced populations in the past. The program BOTTLENECK BOTTLENECK program The past. the in populations influenced have est . 1999) was used to detect evidence of “recent” bottleneck events. events. bottleneck “recent” of evidence detect to used was 1999) .
a
values wasalso out carried values in 0.05 level. a nalysis of
analysis
D h sfwr MPVl Gra & (Garza M_P_Val software the
values (Jost2008).Computations for
The . Thebest m / drift andmigration/drift ARLEQUIN localities localities olecular olecular explanation oftheseprogram settings. mitochondrial geneticdifferentiation among the the the enecks in our data was also tested through the the through tested also was data our in enecks STRUCTURE
after running the programme n+2times, K v the
included in included in the clusters. ariance v.3.1 (Excoffier for each cluster wasselected followingfor eachcluster the F software missing in due to some data loci) nts (e.g. bottlenecks or restriction in gene restrictionin or (e.g. bottlenecks nts ST ST GENAL CR
estimates. (AMOVA) with9
analysis analysis
and EX
ND2
equilibria v.6.5 using99permutations
et alet
( ilasn 2001). Williamson s q ). For microsatellite). For ed by > F 0.5) . 2005) 9,999 with ST -
rf disequilibrium) drift
were performed in F were , 999 permutations. ST
(Weir & with analysed
selection
e the See M -
This article isThis article by protected copyright. All rightsreserved. different geneticdistances ( geographical distance (ln[km]) populationsprev the among permutations, tocompare matricesof distance genetic expressed as implemented in test gene ofscenario isolationby dispersal limitation or alineardistance (IBD),weexpect increaseof estimate the betweencorrelation matrices ofgenetic andgeographic distances. Correlation structureof genetic with geography and analysed withTRACERv.1.6 (Rambaut of toverifyoftheresults.Convergence values wasmonitored theconsistency MCMC chains (Δ 2 ×10 offive2003). Theanalysis consisted independent of2 runs × 10 equilibrium methodimplemented Bayesian v.3.0.4 (Wilson BAYESASS in andRannala, The magnitude and ofgeneflow Estimation iterations after ef the that assumes method this probabilities, (Ciofi 2MOD program the with model obtained alone” “drift and flow/drift” “gene of likelihoods relative the of comparison and
AcceptedM Article
= 0.1; = 0.1; Δ
t ic distance distance withgeographic population 7
To migration from deviations for test The iteration were executed, with
identify identify driverspotential ofpopulation genetic structure, we used F
= 0.5; Δ = 0.5;
s) anda sampling frequencyof1000.Themixing parameterswere adjusted divergence
direction of contemporary gene using flowwas assessed anon the the A
= 0.3) and replicate runs andreplicate performed= 0.3) were withdifferentstarting ISOLDE F
(Ciofi ST
the firstthe
and D extension in et al et est
100,000
for microsatellites,for and . 1999). Five independent runs independent Five 1999). . .
Isolation byd GENEPOP et et al. – t al et drift eq drift steps et f irstlie uain i negligible is mutations microsatellite of fect
2014). . 1999). In the estimation of relative relative of estimation the In 1999). .
discarded asburn
(Rousset 1997) (Rousset uilibrium was based on the estimations the on based was uilibrium istance wasanalysed
ious characteri F 8 ST
iterations aburn (with
for mitochondrial for F , ST
with - /(1 in. of - s F
ed. 1,000,000 MCMC MCMC 1,000,000 10 ST Mantel to tests
) and ) and , using a using Mantel M 000 Under a atrices of -
genes - - seed seed in of ) s
This article isThis article by protected copyright. All rightsreserved. Accepted modelphase (TPM) infinite allele model ( (IAM) contraction events in (pairwise estimates forP4toP15) moderate (P1, groups P2 andP3) (r=0.772;P<0.001),correlated remains similarclusters the Bayesianbetween three approaches (L13, clustersdifferentiated inPortugal twopopulation (L1 andL2 Article withobtained populations ( populations microsatellite markers populations across Genetic Results wasassessedusingof Mantel tests the 100,000permutations. were compared
L18 andL20 The r All pairwise The
structure . microsatellite dataset marked revealed levelsofstructure among genetic chough esults of analysis with the The K G )
, but ENELAND using using
programs differed , demographic events - the the L25) ( F or the some ST PASSAGE Iberia revealed revealed the stepwise the mode stepwise mutation . and main
( Fig. K
populations (P1, P6 (P1, populations =7), =7), D n Peninsula T partitions were very consistent( werevery partitions
est able 1
with v.2.0.11.6 (Rosenberg &Anderson v.2.0.11.6 (Rosenberg a ). The clusters remaining slightly different genetic are
estimates were estimates were BAPS consistent and
( S T slightly slightly the 2 able able ). Noevidence ofbottleneck
to high to high (
BOTTLENECK
K and gene flow highest values among (see Supplementary(see Resul . =9) and =9) first ofthe level The three clustering methods S 1 in their estimate ) . differentiation among Spanish populations
- P9 andP12 highly marked ofstructure among degree chough l (SMM),
suggested historicalsuggested population estimated from significant significant (P<0.001)
- P15) only when P15) only s -
although L3) andthree clusters in Spain of the Fig. the three Portuguesepopulationthe S
ts), yet thenumber of s
1 TRUCTURE
2011) significanceand the was ). optimal
microsatellite loci The clusters genetic
significant values for based on based
found the using
number ofnumber assuming an and highly
( K
=8) clearly clearly =8)
two - This article isThis article by protected copyright. All rightsreserved. Portugal, Spain and 2 bymost shared by 25 control the region (512bp)amplified in577choughsamples, yielded differentiation, in particularly centralIberia. marked basedonmicrosatellites, that than but structureGenetic generations. lowlevels ofimmigrationveryvery andrevealing breeding local migration among rates chough Iberian populations exceed never 0.03(Fig. S1),indicating ofproportion low,generally supported). that indicated and P13)( the (5000) results when pre most consistent, indicating contractions probable population (values ofM polymorphic sitespolymorphic Genetic structure Genetic among pop Migration rates ( representing - bottleneck N T able individuals all with a global with a populations S Iberian are choughpopulations in migration and coalescence using analysis 2 did ). shared ancestral polymorphism The for test migration not bottleneck support a recent (P4 event in sixpopulations e m values of50and500were values haplotype H13haplotype were ( T of migrant of origin ineach ) among clusters choughgenetic able S able using T and form mean able obtained (P1) foronepopulation 4 SMM ( S ) value of0.017( value . 2 ulations onmtDNA based markers The two ). ed The is restricted tocentralis restricted starlike phylogenie starlike T able able - third approach drift equilibrium implemented the in Concatenated sequencesof highest also S 2 ). Themode mtDNA ranging fromranging 0.003to0.240 population reveal tested ( . H . - frequency aplotype H12 is restricted restricted to central H12is aplotype ed , markers T calculated withB based on s haplotype inthe network ( able a considerable - - drift equilibriumdrift (100% Spain (Table S (Table shift test shift test using haplotypes ( S recruitment recent over 2 ). For N ). For higher ( an Fig. TPM 36 haplotypes defined 3 M was expectedly was expectedly less did ) ND2 . - 2 The meanof values , ratio testratio ). degree and three not support any not support H1 and H2) AYESASS Pairwise (680 bp) and (680 bp) ) for the 2 - P6, P8,P10 e MOD of , value was F Fig. ST were were the the s This article isThis article by protected copyright. All rightsreserved. ( in P14 valuesforthenumberThe average nucleotide differences of in(global Hd=0.589),low population inP3and S P1(Hd=0.083)andabsent P5 (Table insertions/deletions ( ofconsisting 19parsimo randomlymating populations. S (PpyA1.12) (Table informativeness ofthe meanmarkers with values rangin (Table S and 0.797(Ppy104) and meanand 0.817(Ppy104), the heterozygosity ranged between expected 0.644(Ppy51) 3.908 and5.892(Table S anaverage(Ppy2P7), with of17.3alleles.A different alleles amplified. Thenumber ofalleles locus per rangedfrom 30 9 (Ppy2P16)to diversityGenetic distribution haplotype among 0.05 significant atthe level (Table S values Acceptedaverage Article estimated frommtDNA data ( The 37 mitochondrial haplotypesThe 37mitochondrial identified were defined All microsatellite wereloci polymorphic all in average π=0.0014) (TableS K=1.498) Table S 6 ). A ny informative sites,five singletonvariable sitesandtwo 6 , verage verage ). Mean observedheterozyg). Mean and nucleotide diversity and nucleotide 4 7 6 ). Overall mitochondrial). Overall haplotype moderate diversity was ). Iberian Iberian ). PIC ). PIC supportthe values high diversity genetic and F IS ranged from 79%oftheestimatesranged 0 to0.945,with being 5 values were values tozeroclose ( ). populations A detailed verage values of verage values allelic richness between ranged is characterization ranged from 0.0023inP14 inP3to zero available populations osity ranged between 0.611(Ppy51) g between 0.597 and (Ppy51) 0.755 ranged from inP3 zero to 2.648 in Supplementary Results. Table S Table by 26 polymorphic sites, by 26polymorphic , of with a total with a of 176 mitochondrial 6 ) asexpected in 7 ). This article isThis article by protected copyright. All rightsreserved. significant differentiated andisolated for line measures population genetic of Drivers structure them (n= (Fig. mo long distance movements ( w variabduring foraging substrates showingthatbirds data nucleipopulation (Table 2). movements movementsindividual 50 than km longer hundredseveral kilometres frequently recorded outside natal their areas,performing long theallowed multiple re ringed in twelve population nuclei ofcapacity choughsamong population nuclei movements of Dispersal marked indivi Acceptedh Article ere samplingere effort wasrelatively low ( vements in central Spainvements simply incentral recordingeffort in because the wasmuch greater this area S2 The correlatio Monitoring movementsof spatial F ). ST 124 was weak was Breeding recruitment eventsof banded individuals as but nestlings were rare, of all correlation explained only of 6.81% the were le timele periods ) occurred in ) occurred longer 300km than in agricultural andpasture land, in andcongregate communal roost andmarginally significant n was - between between from different sighting ofmany ringed individuals ( populations P1andP2( Fig their natal their nucleipopulation (T found between ( Movement data fromMovement data marked isconsistent individuals with our Fig throughout t s . s geographical distances 3 . 3 and over and and involved and involved duals S2 Iberian Iberian the the last ( 30 years S2 variation , Table2 he year ( of individuals of individuals were T geographical geographical , Table2 able able ( regions feeding form mixed flocks onvariable Fig. (r=0.266, recorded 2 GB, ). ), we were able to detect), wewereableto ( Fig. Fig. 135 3 ) Wemoreaccumulated on data have . ). About nine thousand). 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All rightsreserved. individual’s lifetime (GB, unpublished data). natal areas, in engaging retu across Iberia. fromindividuals nuclei different long occasional frequentlyare described as sedentary degree of unusual identified nucleipopulation Lindenmayer genetic like drift sizepopulation in general clusteringapproaches.different Bayesian G flow is betweencorrelation geneticdifferentiation geographic and distances Th among populations ofred Iberian Discussion enetic structure enetic consisted of Accepted Articleese rning tonever natalareas,but permanentlytheir dispersing toregions otherthantheir - term result molecularOur datarevealed not as many as field field ly clearly differentiated fromtheremaining nuclei, asexpectedfrom small their strongly s long are Indeed, Indeed, 2007 study study thatclearly shows longdistancemovements arefrequent, and (Farinha 1991; , striking the of given absence major barriers geographic and - inbreeding, found Iberacross the genetic differentiationat distance movements ( ). influenced by In spite ofourIn spite 15 different non wandering - breeding individualsseem anomadic toshow regularly existence, demographic stochasticitydemographic and/ at least at Blanco 2004) often - billed choughs evidencebilled choughs ofhighdespite capacity. dispersal genetic units the drivers movements canoccur that multipletimes an during extensive but still marked structure genetic and engaging inmostly seven seven shared commonshared i an Peninsula Bullock T he most local major . typically acting in T in hus et al a an area under580,000 an area km nd regional scales genetic clusters clearly clusters genetic , they geographically , . 1983;Cramp 1988; the the ofgeneticanalysis structure partial foraging areas and roosting andforaging areas roosting sites exhibit or reduced geneor reduced flow( sampling extant of the local, altitudinal, and local, and highly geneflow restricted wild high susceptibility to factors isolated populationswere . , Even thoughchoughs animal and suggest supported by three Moore 2006 Moore 2 populations. , revealing an the the weak that gene Fischer & Fischer that ), our ), our This article isThis article by protected copyright. All rightsreserved. Accepted ecological of knowledge, cases frequency anddistance fornegligible consideringtheir choughs highdispersal capacity, populations ( barriers and featureslandscape/ecological most peripheral and isolated Iberianwhen all populations considered were Theobserved. correlation geographic between distance and genetic differentiation was weak structure is notconsistent withthe crossingthe ofmajor barrierssuch as geographical mountains. high basis,includingdaily round ofseveral trips hundredkilometres week withina choughs Articleand breeding season. individualsoutside the breeding or altitudinal patterns and both involve non tobeappear lowinchoughs differences atl variables assessment ofhabitat type qualitative, is more quantitative detailed analysis ofenvironmental to determine ifsmall contribute to genetic the structu local specific may conditions that theirperformance reduce may elsewhere,andthis (Bontenatal areas Local a Local were recorded barriers have in geneflow restricting amongpopulations ocal scalesbeneededocal will definitively to ruleouttheinfluenceof habitat Storfer Storfer daptation may fitness have c et al. et et al such differences between pop throughout between the regions considered for 2012), not . 2007;Sexton extreme structureintheabsence ofgeographic genetic . 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All rightsreserved. Foote ( structure genetic of patterns complex and differentiation on geneflowbetweenprimates and populations markedcetacean canleadto population moreare todocument, difficult isevidencethat butthere Tishko Tishkoff similar case differentiationof genetic wasreported forhumans( social structured Palearctic coinciding with ancestral al. ( Caledonian crows NutcrackerClark’s ( higher than reportedforvalues corvidwith species low population postglacial from colonization latitudes southern reporteddiversity forchough in populations the Isles, British whichare the of result eventsby demographic suchaspopulation bottlenecks. sharp Thisisin contrast with populationbetween of levels as expected ( Accepted Article 2012 et alet ff The extreme level of The extreme level T ) he genetic diversitygenetic et al et al. et haplotypes . vertebrate . 2016 s The recent coalescenceofIberianhaplotypesand the distribution widespread of most p (Wenzel Frankham . 2009;Ross 2009) t ). he organization with different eripheral and eripheral Geographical and Geographical Corvus moneduloides concomitant r nuclei , indicates ayoung postglacial originfor current variation, the genetic s Nucifraga columbiana , and et al et ( Gómez Lunt & 1996; Willi . 2012). . was et al et were This genetic structure attributed tosocio . 2013 suggests that isolated isolated caused byrestricted geneflow and among not populations, ole ofthe Ove et al et ). ecological barriers ecological to dispersal ral 2007; Ferrero 2007; Ferrero S . 2007 ocio population l ) (Abdelkrim nuclear genetic diversity ofIb nuclear diversity genetic ), and fine Iberian PeninsulaIberian social - the the cultural features in in the in the red ) , yet accentuated - differentiation cultural factors ( cultural factors and behavio nuclei most et alet - scale structuregenetic et et - genetic billed choughbilled . 2011; Abellán&Svenning al. the the ofculturalinheritance influence Iberian populationsIberian Kopp show 2012 by as a u differen the the et al et ed and Rosenberg ral non ; Ségurel major Dohms reduced genetic diversity isolation identities. Themost structure patternsstructure . 2014; Kopps 2014; . capacity - human animal suggests suggests tiation patterns erian cho refug et alet et al. et et al et , of , such as rivers, the Newthe show high ium . 2008; small a strongly . 2002; 2013; Rutz u for ghs isalso et al et species species the low the 2014 , the , the . 2014 . ). et et , ; This article isThis article by protected copyright. All rightsreserved. Accepteddistribution range 1994 canphilopatry geneticdifferentiation contribute to ( disp melanorhamphos influence ( features influence 2014 Blanco hierarchical 1997 Article of a versatilerepertoire complex few ( kilometres likely association oftooluseinCaledonian Crows fine with learning have conducted been ( geographical (reviewed barriers in However, cetaceans andmountains habitat anthropogenic fragmentation forprimates, or continental masses for ersal capacity, ), but it explain), but fails to the ) ; ; variabl and D are Corvids known et alet ispersal patternsofhighlyispersal social social Andrews , fine genetic differentiation ofthe different nucleipopulation in m in can differentiation influence geneflowandgenetic betweenpopulations patterns complex , social and . 1997;Blanco & e - scale spatial geneticstructure whitein the unrelated ost cases nesting and familiar interactions Abdelkrim (Beck of the Iberian 2014) recruitment tohappen appears natal onlyat areas vocalizations ( systems ritualized , foraging et al et . Restricted due. Restricted socialhasdispersal barriers to beenreported the complexthe structure genetic cannot beexplained for et al et . 2008). Tella 1999 their hightheir cognitive capacityand studiesseveral on social Clayton &Emery and communal roosting and communal roosting lack lack of . 2012; chough habits interactions amonginteractions groupmembers Andrews Laiolo In contrast, In contrast, , isolation isolation by distance . Genetic . Genetic largestructure scalesinat species bird with Rutz requiring learning from conspecifics ) (Bignal (Bignal ; species arefrequently by influenced et al high 2014). et alet . 2000 mate et al while . 2012 2007 Nyholm1986; Weatherhead & Forbes , 2001 . and nest 1997 ). Iberian patterns ). Previous studies have reported a Choughs ), ; - within th within scale geneticscale across structure a Blanco & all of all - site fidelity ( red involving complex - - by cultural billed choughs winged chough, which may also showalso . This e ( Still Still Tella 1999 continuous level of Banda & et alet simply by ( evolution remarkabl Bignal potentially potentially socio . 1987; ), show strict natal strict Corcorax including - Blanco, cultural et al to . y high . . This article isThis article by protected copyright. All rightsreserved. non choughs knowledge duesolelyto be high dispersalcapa and interact withregularly O Conclusion and pairing explainsocial to barriers” evolutionary divergence 2014 features( with pronounced social and culturally evolutionary inherited can changes drive 2009; Ross important also are practises 2014). Furthermore, factors cultural dispersal habitat types social and/or features, group structure such as strong stability andsex barriers ecological regionalat scale choughs is small 2015 by isolation distance ( moderate Acceptedur long Article - human ; ) , anda populations Foote , , - - is unique among birds, andis uniqueamong birds, only comparableto term studydemonstratesth to can , t unlike et al et recruitment primates - et al et high he complex genetic lack of lack explain . 2013 to patterns ofisolation bydistance,oradaptation. colonization To . 2016 dispersal capacity significantlyis often associated any case ( city Agudo isolation bydistance isolation , and cetaceans. Our findings Agudo the structure pronounced observed genetic inseveral species ( ) , the ). . , T deserv In lightofthis evidence, here is here et al et individuals of differentpopulation nuclei reported forreported birds marked structure observed genetic isunprecedented, and factors regulating factors et al et . 2011 ing of structure also Langergraber . 2011; Mira s . Natal like at choughs in at engage regular long ). future research increasing evidence evidence increasing that The extreme ofgeneticstructure ofIberian level ethnolinguistic boundaries is , described - range generally that un are gene flow in human populations ( populations human in flow gene et et al et al philopatry orphilopatry toparticular philopatry suggest thatsuggest we propose we propose , . 2013; . 2011;Rutz partly partly . here for here firstthe time due to due to genetic wild differentiation in species constrained bygeographical or that induced bynon Graciá highly fragmentedhighly andisolated reported forreported humans, a a combination of genetically a combination of genetically complex pattern mec et alet et al and traditional social h with across Iberia . 2012; Kopps anism anism . 2015; - distance movements a pattern of a pattern - random of patterns for the of “isolation by of “isolation Pellegrino s T of . ishkoff Iberian our Given this - some social biased et et al Andrews unlikely . wild et al et et al et . . This article isThis article by protected copyright. All rightsreserved. Biological Journalof the Society Linnean condition, investment onsize, and immunocompetence in nestlingred Influence Banda E,Blanco G(2008) of hatching asynchrony and within moleculardetect adaptation basedon a Fst Antao T,LopesA,RJ,Beja Yamagiwa(eds J,Karczmarski pp.289 L), andPrimates Research Cetaceans: Field andConservationof Com Andrews K(2014)Population genetics the in conservation of cetaceansandprimates. In: Distributions continental a Agudo R,C,Hiraldo (2011)Evidence F,DonázarJÁ ofconnectivity Rico between of Journal the Linnean Society divergence arelinkedto QuaternaryLate climate the Peninsula. stabilityin Iberian (2014) P,SvenningJC Abellán Refugiawithin refugia ofthecolonisation history tool Abdelkrim genetic J,HuntGR,GrayRD,GemmellPopulation structureand NJ(2012) R CGL2010 and the UID/AGR/04033/2013project Funds forNational byPortugueseFoundation ScienceandTechnology and Internacionalization Programme, ProjectPOCI under by Florestas Conservação daNatureza edas for collaborationGonçalves their infieldworkand sampleand collection, J.A.Ribeiro, Sanchéz Gomes, Cuevas, C.Dionísio,J.A. Fargallo, M.J. Fernandes, A.Frazão,J.M.García, F.Gómez, C. F.Barros, Á.Barros, P.Barros,R. Banda, S.Barbosa, Bastos, that improved manuscript. the theeditorandfiveanonymousP. Laiolo, subject reviewers provided constructive comments Acknowledgements of reviewing isalsoThere including identify interactions eferences Accepted Articleisolation European Investment FundsbyFEDER/COMPETE/POC POPH projects projects 082/2002, BOS2003 ing P. Laiolo, P. Laiolo, - / the the main FSE 15726, CGL2015 15726, the proximate by may nd differentiatedinsular populationsinahighlymobile species. , a potential potential 17, 1 social need to expand knowledgeneed to our program(Programa Operacional Humano/FundoEuropeu)Potencial Social be responsible be responsible R. López, Pérez J.Mouriño,L.B.Pais,J.M. F.Martínez, – hypothes 12. barriers - Zapata, M.Santos, N.Santos,F.Silva, J.L existence of mechanisms stronggenetic this structure driving - e 66381 must considered be also s We F.Álamo,R. Andrade,B.Arroyo,A.Artázcoz, E. thank , - and the PhDGrant SFRH/BD/77872/ using New Crow. Caledonian o 113 for n - 05066, CGL2009 - genetic structuring of wildpopulations Pereira A,LuikartG(2008)LOSITAN:Aworkbench to - , 13 social social genetic differentiation P byregional andnational Spanish – 28. , group - for technical assistance. - 94 outlier method. 308. Springer, Japan.308. Springer, , 675 on other on other identit – - 684. 12753 . y - highly patterns i and - . 01 - F BMC Bioinformatics BMC C02 orthcoming a approaches I - within – 0145 PLoS ONE complex societies vertebrate M. Opera - 01/BOS, PPIC10 . Tella, . Tella, I. Torre andH. Vale n endemism andgenetic C. Blanco, - This work wassupported plex plex Mammalian Societies FEDER - group 2011 t - ional Competitiveness billed choughs. (FCT) g - , brood parental overnments. , Instituto da Instituto 7 financed through where phenomena reproducti are needed, , e36608. - - 006958 and García, A. Diversity and J. Blasco, J.A. , through the , 9 - , 0094 Biological 323. ve imed - skew 3036, , , at - . 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All rightsreserved. Accepted4015 L(2008)Jost GSTandits r multilocus genotype data. OR,Jones Wang COLONY: J(2010) aprogram for sibshipfrom and inference parentage 1332. assistawith the structure Hubisz MJ, D,StephensM,PritchardJK (2009) Inferringweakpopulation Falush genetics. G, Guillot F,EstoupMortier A(2005) package GENELAND: forlandscape acomputer Genetics. G,Estoup F,CossonGuillot A,Mortier Statistical JF(2005)A ModelforLandscape Spatial Peninsula. Iberian the thelast Eagletop predator bottlenecksandduring century: expansions oftheEurasian Owl in E,OrtegoGraciá J,GodoyJA(2015)signatures ofdemographicGenetic an changesin N), Ferrand pp. 155 in the Iberian Peninsula. In: refugia:Gómez patter DH(2007)Refugiawithin A,Lunt Royaltransactions ofthe ofLondon.Society Seriessciences B,Biological H(2011)Gintis Gene microsatellite loci. 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Journal BiologyJournal of Avian , 103 Evolution – – - 106. 5999. locus study. s , 28 - , , 2537 specific structur genetic 68 , , 2 - , 1 , 229 scale population scale 99 – – 15. 2539. PLoS Genetics , 313 – - 232. The Journal of statistics under Orcinus orca – , 320. 46 Science , 462 – , , 475. 4 e 236 , ). , , This article isThis article by protected copyright. All rightsreserved. The authors noconflictsdeclare ofinterest. Accepted ofConflicts interest authors read andapprovedsubmitted version. the contributed to th J.A.D.analyses performed laboratory the bulkG.B. the conducted of long the F.M., J study; the F.M., J.A.D.,B.M.andG.B.designed J.A.C., andG.B. E.B. thestudy; coordinated Author contributions KX024343GenBank (accessions (doi: repository The Data Accessibility Articleevolution and adaptation. Zhang G,LiC, Q Wright Isolation S(1943) bydistance. genotypes. WilsonRannala B(2003)Bayesian inference GA, ofrecent multilocusmigration using rates 542. populations is exacerbated bydriftand selection. WilliJ, Buskirk Y,Van SchmidM (2007) B,Fischer Genetic ofisolation fragmented Genetics Conserervation genetic WebsterWenzel LMI, MA, Blanco G Structure. Weir BS,Cockerham (1984)Estimating CC F ecological in Weatherhead philopatry PJ,ForbesMRL Natal birds: inpasserine (1994) or genetic Ecology Notes software foridentifying andcorrecting in genotyping errors microsatellite data. Van Oosterhout WF, C,Hutchinson W Phylogenetics andEvolution Phylogeography boreal of bird,gray awidespread jay the ( C,KlickaVan ElsP,Cicero J(2012) highgeneticdiversity: Highlatitudes and field .A.C., O.F., J.L.G., P.T., D.C. and G.B. collected biological .A.C., O.F.,J.L.G.,P.T.,D.C. G.B.biological and samples; collected O.F.,J.L.G.,and diversity diversity in European red monitoring data and Evolution Genetics fluences? fluences? , 4 10.5061/dryad.684v0 e interpretation of the results; , 535 , , et 38 163 – Behavioral Behavioral Ecology 538. al. , 1358. , , 1177 Science 13 (2014) Comparative(2014) genomics insights intoaviangenome reveals , 1213 , 63 microsatellite – , 456 1191. - - , KX024378 and billed chough( 346 – - 1230. term bandingandmonitoring; population F.M.and – ) . Mitochondrial from DNAsequences are available et al. et , 1311 465. ills DP, Shipley P(2004) MICRO DP,Shipley ills Genetics ; F.M., J.A.D.,and G.B. B.M.analysed the , (2012) Pronouncedstructureandlow genetic 5, 426 allele – 1320. - F.M., B.M.andG.B.manuscript. wrotethe All Statistics for Statistics theAnalysis of Population , 28 Pyrrhocorax Pyrrhocorax pyrrhocorax Jo KX024396 – dataset , 114 433. urnal of urnal Evolutionary Biology - 138. s Perisoreus canadensis have been - KX024431). archived in the Dryad - CHECKER: ) populations. Molecular Molecular ). , 20 Molecular Molecular data and , 534 – This article isThis article by protected copyright. All rightsreserved. d c b a Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Spain Portugal Portugal Portugal Portugal shown inparentheses. H onindividualgenotypesbased frommicrosatellite loci. Table 1 Tables andFigures Country Number of samples with successful PCR amplification for mitochondrial DNA and micr mitochondrial DNA and for amplification PCR successful with samples Number of (A) Aproximate minimum and maximum altitudes of the areas used by each population. each by used theareas maximum of minimumand altitudes Aproximate ( A O Accepted) Article Crevices and caves (sea cliff); (B) crevice and caves (inland); (C) human (C) (inland); caves and crevice (B) cliff); (seacaves and Crevices Grassland ; (B) agricultural areas (cereal crops); (C) short scrubland. short (C) (cereal crops); areas ; agricultural (B) Grassland - observed heterozygosity, Málaga Granada Murcia Albacete Teruel Monegros Los Urbasa Asturias Morte da Costa Riodeporcos Vilarín Lastra da Enciña Barroso Estrela da Serra Mós de Porto Sagres Area TOTAL east Madrid west Madrid Segovia Azálvaro Lobos Río Guadalajara Castuera Hornachos Real Ciudad Sampling localities, Locality L25 L24 L23 L22 L21 L20 L19 L18 L17 L16 L15 L14 L13 L12 L11 L10 L9 L8 L7 L6 L5 L4 L3 L2 L1 Dominat Dominat habitat A,B,C A,B,C A,B,C A,B,C A,B,C A,C A,C A,C A,C A,C A,C A,C B,C B,C B,C B,C B,C B,C B,C B,C B,C B,C A A A habitat variables, H a E - sites Nest B,C B,C B,C B,C B,C B,C B,C expected heterozygosity,expected with standard (SD) deviation A A C B B B B B C C B B B B B B B B b Altitude Altitude range 0 0 1000 1200 1000 1000 1000 1000 2000 1300 2000 1400 1600 1200 1200 2000 1200 1200 700 500 600 500 200 600 500 200 600 500 900 300 400 500 500 300 500 200 600 600 900 900 800 700 600 900 700 800 700 500 - - 200 200 ------ - - - - - c sample andgenetic sizes, indicesdiversity Genetic Genetic cluster P P P P P P P P P P P P P P P P P P P P P P 15 15 14 14 14 13 ? 12 ? 11 10 10 ? 7 6 5 4 4 4 3 2 2 1 9 8 - made structures. (mtDNA) N Sample A size 577 37 40 14 16 19 10 13 25 24 56 57 17 23 15 14 17 15 35 23 24 24 35 6 9 9 – osatellites without full without osatellites mean number locus, per allele d (microsat) Sample size 590 39 38 14 16 19 10 13 25 24 57 56 18 23 15 17 17 42 23 24 14 24 38 6 9 9 d (0.98) (0.99) (0.55) (0.65) (0.54) (0.52) (0.40) (0.57) (0.38) (0.48) (0.81) (1.15) (0.65) (0.60) (0.49) (0.50) (0.64) (0.51) (0.48) (1.22) (0.68) (0.80) (0.55) (0.79) (0.76) 10.60 10.60 10.30 10.30 - 9.70 9.70 9.30 5.10 7.30 7.00 6.00 3.60 5.10 5.10 4.50 9.80 9.80 7.70 8.30 7.20 8.40 6.90 6.20 5.60 9.00 9.80 5.80 9.80 sibs. N A (0.02) (0.03) (0.04) (0.03) (0.05) (0.07) (0.09) (0.07) (0.06) (0.05) (0.04) (0.05) (0.02) (0.04) (0.02) (0.03) (0.04) (0.04) (0.05) (0.04) (0.04) (0.03) (0.04) (0.04) (0.05) 0.75 0.75 0.74 0.65 0.71 0.74 0.76 0.55 0.54 0.63 0.62 0.80 0.80 0.74 0.82 0.80 0.80 0.85 0.81 0.72 0.81 0.72 0.85 0.80 0.68 0.73 0.74 H O (0.03) (0.02) (0.04) (0.04) (0.03) (0.03) (0.05) (0.06) (0.04) (0.04) (0.03) (0.03) (0.02) (0.02) (0.02) (0.02) (0.02) (0.03) (0.04) (0.04) (0.03) (0.02) (0.02) (0.02) (0.04) 0.79 0.79 0.79 0.63 0.73 0.74 0.72 0.57 0.59 0.62 0.60 0.78 0.78 0.77 0.78 0.79 0.75 0.80 0.79 0.70 0.72 0.75 0.77 0.81 0.70 0.79 0.79 H E This article isThis article by protected copyright. All rightsreserved. 9862 localities c natal area. b a TOTAL Navarra and L25) Madrid (L24 Segovia (L23) (L22) Azálvaro (L17) CiudadReal Granada(L15) Murcia (L14) Teruel (L12) (L11) MonegrosLos Barroso (L4) (L2) Porto demós Location Peninsula. Iberian the Table 2 Frequencies of movements longerthan 50 km from thenatal areasto Population recentlymonitored and notincludedin the genetic analysis. AcceptedNesting recruitedas breeders intheir natal areas; noindividual was found recruited outside its Article ). a from asubsample of the records for whichthe distance of movements calculated was ( Results ofthemarkResults nestli 5025 3322 127 915 154 326 100 ngs 19 21 29 9 3 individuals Marked grown adults fully 3964 3133 117 602 49 16 45 1 0 1 0 0 - - recapture andfield monitoring of 12choughpopulation nuclei in Recaptured/res indivi duals 2753 1715 734 143 76 39 27 9 3 1 3 3 ighted recor 4242 1025 5269 480 799 98 30 55 ds 4 2 3 7 3 Nestli recrui ted 124 ngs 93 23 6 2 ------ b 50 Number of movement - 300/>300 km (mean 1747/210 443/18 distance recapture/resight 179 1041/2 (9 49/1 (12 21/1 (177) 10/0 0 2/0 0 1/0 records 1/0 /4 (4 /6 /8 (10 (126) (42 (2 (18 8 (17 (8 10 70 , 4 ( 4 km 3 3 ) 198 c 8 ) ) ) 9 ) ) ) 6 ) ) ) ) ing period Monit 2011 1993 2011 2014 2014 2011 1985 2012 2012 1985 2014 1988 oring 2016 2015 2015 2015 2015 2016 2014 2015 2016 2016 2016 2016 n = ------ This article isThis article by protected copyright. All rightsreserved. Accepted different population nuclei. recorded forthree individual choughs ( shown. SeeFig. movements ofthesameindividuals.or different movements Only 50km than longer are by different tothecolours), resighting/recapture Each sing site. line represents nuclei.population Lines represent individual movements population nuclei (indicated from Fig. 3 Article frequencies. byseparated asinglenucleotide change, proportional to andcirclesizes are haplotype highlighting sixmostwith the commonhaplotypes differentcolours; all haplotypes are tosampleproportional size(see Table1).Thenetwork includ corresponding to in colours those the haplotypeofeachpiechartis Thesize network. mtDNA Pie inthecharts map sequences. thefrequencyofwith haplotype, indicate each Fig. 2 STRUCTURE estimatedclusters program byeach (K=7for colours represent posteriorthe probability of assignment using clusters in ( with circle size proportional tosample size,see 1).ColoursTable correspond red Fig. 1 legendsFigure - billed chough range(grey distribution area)andsampling (numbered localities circles, GENELAND Geographical distributionGeographical ( Genetic structure among populations ofthered Iberian Dispersal capacityDispersal of red Iberian ). b ). ( b S2 , ) Results ofgenetic analysis structureinferredfrom the microsatellite data BAPS for the exact offor theexact location individual ( records. and and STRUCTURE a ) andmedian band - . Vertical correspondbars toindividual choughs and billed choughs.( numbers 1N4,H1L and GENELAND - joining networkjoining ( , K=9 for , K=9 a to eachof anoptimal number of ) Dispersal range of12chough - es the 37haplotypes es the identified, billed chough. ( b ) Examplesmovements of BAPS b ) ofthe concatenated 6FR) from three , and K=8for , and to genetic to genetic a le orle multiple ) Map ) Map of the This article isThis article by protected copyright. All rightsreserved. Accepted Article This article isThis article by protected copyright. All rightsreserved. Accepted Article